• Title/Summary/Keyword: 2b-site

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Analysis of Transcripts Expressed from the UL47 Gene of Human Cytomegalovirus

  • Hyun, Jong-Jun;Park, Hyo-Soon;Kim, Ki-Ho;Kim, Hung-Jin
    • Archives of Pharmacal Research
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    • v.22 no.6
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    • pp.542-548
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    • 1999
  • The UL47 gene (b 60390-b 60388) located in the unique long region of the human cytomegalovirus (HCMV) AD169 strain genome was analyzed RNA mapping. Northern blot analysis showed that the UL47 gene was expressed at late times after infection (72 h postinfection). The 9.7-kb transcript was expressed in the infected cells but not in phosphonoformate-treated cells at 72 hpi, indicating that the UL47 gene was only expressed at late times after infection. To map the 5'-end and 3'-end of UL47 transcripts, primer at late times after infection. To map the 5'-end and 3'-end of UL47 transcripts, primer extension and RNase protection analysis were performed. Primer extension analysis revealed that the transcription initiation site of UL47 was located in 27 bp downstream (b 60323) of the TATA box motif. The sizes of UL47 ORF (approximately 2.9-kb) and UL48 ORF (approximately 6.7-kb) deduced from computer sequence analysis suggest that the expressed 9.7-kb transcript of UL47 uses the 3'-end polyadenylation signal of Ul48. The result of RNase protection determined that the 3'-end of UL47 RNA utilized the 3'-end polyadenylation signal of UL48, which is located in HCMV genome b 70082.

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Molecular Orbital Anaysis of Water Activation on TiO2(110) Surface (TiO2(110) 표면에 흡착된 물분자의 결합 활성화에 관한 MO 연구)

  • Kang, Dae-Bok;
    • Journal of the Korean Chemical Society
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    • v.46 no.3
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    • pp.179-186
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    • 2002
  • A molecular orbital analysis based on the extended Huckel calculations has been carried out to study the OH bond activation of water on the $TiO_2$(110) surface. $H_2O$ binds with its axis perpendicular to the surfac on top of the five-coordinate $Ti^{4+}$ atom via its $3a_1$ orbital. In this bonding situation, the two-coordinated bridging $O^{2-}$ atom ($O_b$, basic site) on $TiO_2$(110) is too distant from an H atom of water to form hydrogen-bondig interactions with water that facilitate O-H bond cleavage. It has been elucidated that the O-H bond is appreciably weakened when the water molecule is tilted to give a hydrogen bond with the $O_b$ atom. This mechanism includes mutual transfer of electron density from the $3a_1$ orbital of the water molecule to the $Ti^{4+} 3d_{z2}$ orbital and from the $O_b$ P orbitals to the $2b_1$ of the adsorbed water molecule This should result in lengthening of the O-H bond in the surface complex and the subsequent dissociation into the fragments OH and H.

Phenology and Reproductive Effort of Two Zostera marina L. Populations on the Southern Coast of Korea (남해 연안에 생육하는 거머리말(Zostera marina L.) 개체군의 생물계절학과 생식 능력)

  • Lee, Sang-Yong;Lee, Sung-Mi;Choi, Chung-Il
    • Ocean and Polar Research
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    • v.27 no.1
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    • pp.67-74
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    • 2005
  • The phonology and reproductive effort of Zostera marina L. (Dongdae Bay and Aenggang Bay) on the southern coast of Korea were investigated from September 2002 to August 2003. The Dongdae Bay(DB) exhibited a finer sediment grain size with richer organic content than the Aenggang Bay(AB). Water temperature at DB site ranged from $7.5^{\circ}C$ in February to $25.5^{\circ}C$ in August, while water temperature at AB site ranged rrom $6.7^{\circ}C$ in February to $25.2^{\circ}C$ in september. Both populations exhibited a perennial lifB span, but the sexual reproductive effort was significantly higher at DB than AB. The reproductive shoots of DB site were developed from February to June 2003, while those of AB site from March to June 2003. At DB site, the frequency of reproductive shoot ranged from 11 to 33%, whereas reproductive shoot of AB did below 5%. Reproductive structures were larger and more abundant at DB, with the exception of number of fruits and seeds per spadix. Water temperature seems most important to regulate the timing of reproductive phase of Z. marina populations for two study sites.

Nucleotide Sequence, Structural Investigation and Homology Modeling Studies of a Ca2+-independent α-amylase with Acidic pH-profile

  • Sajedi, Reza Hassan;Taghdir, Majid;Naderi-Manesh, Hossein;Khajeh, Khosro;Ranjbar, Bijan
    • BMB Reports
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    • v.40 no.3
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    • pp.315-324
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    • 2007
  • The novel $\alpha$-amylase purified from locally isolated strain, Bacillus sp. KR-8104, (KRA) (Enzyme Microb Technol; 2005; 36: 666-671) is active in a wide range of pH. The enzyme maximum activity is at pH 4.0 and it retains 90% of activity at pH 3.5. The irreversible thermoinactivation patterns of KRA and the enzyme activity are not changed in the presence and absence of $Ca^{2+}$ and EDTA. Therefore, KRA acts as a $Ca^{2+}$-independent enzyme. Based on circular dichroism (CD) data from thermal unfolding of the enzyme recorded at 222 nm, addition of $Ca^{2+}$ and EDTA similar to its irreversible thermoinactivation, does not influence the thermal denaturation of the enzyme and its Tm. The amino acid sequence of KRA was obtained from the nucleotide sequencing of PCR products of encoding gene. The deduced amino acid sequence of the enzyme revealed a very high sequence homology to Bacillus amyloliquefaciens (BAA) (85% identity, 90% similarity) and Bacillus licheniformis $\alpha$-amylases (BLA) (81% identity, 88% similarity). To elucidate and understand these characteristics of the $\alpha$-amylase, a model of 3D structure of KRA was constructed using the crystal structure of the mutant of BLA as the platform and refined with a molecular dynamics (MD) simulation program. Interestingly enough, there is only one amino acid substitution for KRA in comparison with BLA and BAA in the region involved in the calcium-binding sites. On the other hand, there are many amino acid differences between BLA and KRA at the interface of A and B domains and around the metal triad and active site area. These alterations could have a role in stabilizing the native structure of the loop in the active site cleft and maintenance and stabilization of the putative metal triad-binding site. The amino acid differences at the active site cleft and around the catalytic residues might affect their pKa values and consequently shift its pH profile. In addition, the intrinsic fluorescence intensity of the enzyme at 350 nm does not show considerable change at pH 3.5-7.0.

On the vibration influence to the running power plant facilities when the foundation excavated of the cautious blasting works. (노천굴착에서 발파진동의 크기를 감량 시키기 위한 정밀파실험식)

  • Huh Ginn
    • Explosives and Blasting
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    • v.9 no.1
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    • pp.3-13
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    • 1991
  • The cautious blasting works had been used with emulsion explosion electric M/S delay caps. Drill depth was from 3m to 6m with Crawler Drill ${\phi}70mm$ on the calcalious sand stone (soft -modelate -semi hard Rock). The total numbers of test blast were 88. Scale distance were induced 15.52-60.32. It was applied to propagation Law in blasting vibration as follows. Propagtion Law in Blasting Vibration $V=K(\frac{D}{W^b})^n$ were V : Peak partical velocity(cm/sec) D : Distance between explosion and recording sites(m) W : Maximum charge per delay-period of eight milliseconds or more (kg) K : Ground transmission constant, empirically determind on the Rocks, Explosive and drilling pattern ets. b : Charge exponents n : Reduced exponents where the quantity $\frac{D}{W^b}$ is known as the scale distance. Above equation is worked by the U.S Bureau of Mines to determine peak particle velocity. The propagation Law can be catagorized in three groups. Cubic root Scaling charge per delay Square root Scaling of charge per delay Site-specific Scaling of charge Per delay Plots of peak particle velocity versus distoance were made on log-log coordinates. The data are grouped by test and P.P.V. The linear grouping of the data permits their representation by an equation of the form ; $V=K(\frac{D}{W^{\frac{1}{3}})^{-n}$ The value of K(41 or 124) and n(1.41 or 1.66) were determined for each set of data by the method of least squores. Statistical tests showed that a common slope, n, could be used for all data of a given components. Charge and reduction exponents carried out by multiple regressional analysis. It's divided into under loom over loom distance because the frequency is verified by the distance from blast site. Empirical equation of cautious blasting vibration is as follows. Over 30m ------- under l00m ${\cdots\cdots\cdots}{\;}41(D/sqrt[2]{W})^{-1.41}{\;}{\cdots\cdots\cdots\cdots\cdots}{\;}A$ Over 100m ${\cdots\cdots\cdots\cdots\cdots}{\;}121(D/sqrt[3]{W})^{-1.66}{\;}{\cdots\cdots\cdots\cdots\cdots}{\;}B$ where ; V is peak particle velocity In cm / sec D is distance in m and W, maximLlm charge weight per day in kg K value on the above equation has to be more specified for further understaring about the effect of explosives, Rock strength. And Drilling pattern on the vibration levels, it is necessary to carry out more tests.

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A Statistical Study of Radiolucent Foreign Bodies in Food and Air Passage (단순 흉부 방사선 검사상 발견되지 않은 식도 및 기도 이물의 임상 통계적 고찰)

  • 손영규;이창업;배광식;박문서;이선철
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1983.05a
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    • pp.9.2-9
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    • 1983
  • We have observed 74 cases of radiolucent foreign bodies in food and air passage in E. N. T. department of Hang Gang Sacred Hospital, from Jan. 1972 to Mar. 1983. The following results were obtained. 1) Distribution of location was 56 cases(75.6 %) in food passage and 18 cases (24.4 %) in air passage. 2) In age distribution, 46 cases (82.1 %) of F. B. in food passage were found in patient over 21 years old and 12 cases (66.7 %) of F. B. in air passage were found in patient under 5 years old. 3) Female seems to be more frequently involved than male in cases of F. B. of food passage and in cases of air passage, male is more frequent. Food passage male: female - - - 25 : 31 Air passage male: female - - - 10 : 8 4) Meats was the most frequent foreign body in food passage (19 cases-33.9%), and the bony pieces was next (17 cases-30.4 %). Plastic material was the most frequent foreign body in air passage (9 cases-49.9 %), and the vegetable seed was the second (4cases-22.0 %). 5) In the location of F. B. in food passage, the first narrowing of the esophagus was the most frequent site and in air passage, the bronchus-especially right bronchus-was the most frequent site. 6) In the duration of lodgement, 44 cases(78.6 %) of F.B. in food passage were visited with - in 24 hours, and 11 cases (61 %) of F. B. in air passage were visited with - in 24 hours. 7) Simple chest PA was checked in all patient and then, esophagogram was checked in 34 cases of F.B. in food passage, among them positive finding was noticed in 23 cases. 8) Removal of F. B. in food passage by esophagoscopy was performed in 54 cases (96.4 %), but in the cases of air passage, removal of F. B. by bronchoscopy was performed in 14 cases (77.8 %). 9) Complications of food passage by the F. B. itself or esophagoscopy were esophageal mucosal laceration (1 cases-1.8 %) and esophageal perforation (1 case -1. 8 %) and complication of air passage by F. B. itself or bronchoscopy were atelectasis (2 cases -11.1 %) and pneumonia (3 cases-16.7 %).

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Enhancement of Lipid Production under Heterotrophic Conditions by Overexpression of an Endogenous bZIP Transcription Factor in Chlorella sp. HS2

  • Lee, Hansol;Shin, Won-Sub;Kim, Young Uk;Jeon, Seungjib;Kim, Minsik;Kang, Nam Kyu;Chang, Yong Keun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.10
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    • pp.1597-1606
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    • 2020
  • Transcription factor engineering to regulate multiple genes has shown promise in the field of microalgae genetic engineering. Here, we report the first use of transcription factor engineering in Chlorella sp. HS2, thought to have potential for producing biofuels and bioproducts. We identified seven endogenous bZIP transcription factors in Chlorella sp. HS2 and named them HSbZIP1 through HSbZIP7. We overexpressed HSbZIP1, a C-type bZIP transcription factor, in Chlorella sp. HS2 with the goal of enhancing lipid production. Phenotype screening under heterotrophic conditions showed that all transformants exhibited increased fatty acid production. In particular, HSbZIP1 37 and 58 showed fatty acid methyl ester (FAME) yields of 859 and 1,052 mg/l, respectively, at day 10 of growth under heterotrophic conditions, and these yields were 74% and 113% higher, respectively, than that of WT. To elucidate the mechanism underlying the improved phenotypes, we identified candidate HSbZIP1-regulated genes via transcription factor binding site analysis. We then selected three genes involved in fatty acid synthesis and investigated mRNA expression levels of the genes by qRT-PCR. The result revealed that the possible HSbZIP1-regulated genes involved in fatty acid synthesis were upregulated in the HSbZIP1 transformants. Taken together, our results demonstrate that HSbZIP1 can be utilized to improve lipid production in Chlorella sp. HS2 under heterotrophic conditions.

e-Pharmacophore modeling and in silico study of CD147 receptor against SARS-CoV-2 drugs

  • Nisha Kumari Pandit;Simranjeet Singh Mann;Anee Mohanty;Sumer Singh Meena
    • Genomics & Informatics
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    • v.21 no.2
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    • pp.17.1-17.12
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    • 2023
  • Coronavirus has left severe health impacts on the human population, globally. Still a significant number of cases are reported daily as no specific medications are available for its effective treatment. The presence of the CD147 receptor (human basigin) on the host cell facilitates the severe acute respiratory disease coronavirus 2 (SARS-CoV-2) infection. Therefore, the drugs that efficiently alter the formation of CD147 and spike protein complex could be the right drug candidate to inhibit the replication of SARS-CoV-2. Hence, an e-Pharmacophore model was developed based on the receptor-ligand cavity of CD147 protein which was further mapped against pre-existing drugs of coronavirus disease treatment. A total of seven drugs were found to be suited as pharmacophores out of 11 drugs screened which was further docked with CD147 protein using CDOCKER of Biovia discovery studio. The active site sphere of the prepared protein was 101.44, 87.84, and 97.17 along with the radius being 15.33 and the root-mean-square deviation value obtained was 0.73 Å. The protein minimization energy was calculated to be -30,328.81547 kcal/mol. The docking results showed ritonavir as the best fit as it demonstrated a higher CDOCKER energy (-57.30) with correspond to CDOCKER interaction energy (-53.38). However, authors further suggest in vitro studies to understand the potential activity of the ritonavir.

Suppression of Human GD3 Synthase (hST8Sia I) Expression Induced by Retinoic Acid in Human Melanoma SK-MEL-2 Cells (흑색종세포주 SK-MEL-2에서 레티노이드에 의한 GD3합성효소(hST8Sia I)의 발현억제)

  • Kwon, Haw-Young;Kang, Nam-Young;Lee, Young-Choon
    • Journal of Life Science
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    • v.20 no.5
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    • pp.655-661
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    • 2010
  • To elucidate the mechanism underlying the suppressive regulation of hST8Sia I expression in retinoic acid (RA)-induced SK-MEL-2 cells, we characterized the promoter region of the hST8Sia I gene. Functional analysis of the 5‘-flanking region of the hST8Sia I gene by the transient expression method showed that the -1146 to -646 region, which contains putative binding sites for transcription factors c-Ets-1, CREB, AP-1 and NF-kB, functions as the RA-repressive promoter in SK-MEL-2 cells. Site-directed mutagenesis and ChIP analyses indicated that the NF-kB binding site at -731 to -722 is crucial for the RA-induced repression of hST8Sia I in SK-MEL-2 cells. In addition, the transcriptional activity of hST8Sia I suppressed by RA in SK-MEL-2 cells was strongly inhibited by extracellular signal-regulated protein kinase (ERK) inhibitor U0126 and protein kinase C (PKC) inhibitor GO6976, as determined by RT-PCR and luciferase assay of hST8Sia I promoter containing the -1146 to -646 regions. These results suggest that RA markedly modulates transcriptional regulation of hST8Sia I gene expression through the PKC/ERK signal pathway in SK-MEL-2 cells.

In-silico Studies of Boerhavia diffusa (Purnarnava) Phytoconstituents as ACE II Inhibitor: Strategies to Combat COVID-19 and Associated Diseases

  • Rahul Maurya;Thirupataiah Boini;Lakshminarayana Misro;Thulasi Radhakrishnan
    • Natural Product Sciences
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    • v.29 no.2
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    • pp.104-112
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    • 2023
  • COVID-19 caused a catastrophe in human health. People infected with COVID-19 also suffer from various clinical illnesses during and after the infection. The Boerhavia diffusa plant is well known for its antihypertensive activity. ACE-II inhibitors and calcium channel blockers are reported as mechanisms for the antihypertensive activity of B. diffusa phytoconstituents. Various studies have said ACE-II is the virus's binding site to attack host cells. COVID-19 treatment commonly employs a variety of synthetic antiviral and steroidal drugs. As a result, other clinical illnesses, such as hypertension and hyperglycemia, emerge as serious complications. Safe and effective drug delivery is a prime objective of the drug development process. COVID-19 is treated with various herbal treatments; however, they are not widely used due to their low potency. Many herbal plants and formulations are used to treat COVID-19 infection, in which B. diffusa is the most widely used plant. The current study relies on discovering active phytoconstituents with ACE-II inhibitory activity in the B. diffusa plant. As a result, it can be used as a treatment option for patients with COVID-19 and related diseases. Different phytoconstituents of the B. diffusa plant were selected from the reported literature. The activity of phytoconstituents against ACE-II proteins has been studied. Molecular docking and ligand-protein interaction computation tools are used in the in-silico experiment. Physicochemical, drug-likeness, water solubility, lipophilicity, and pharmacokinetic parameters are used to evaluate phytoconstituents. Liriodenine has the best drug-likeness, bioactivity, and binding score characteristics among the selected ligands. The in-silico study aims to find the therapeutic potential of B. diffusa phytoconstituents against ACE-II. Targeting ACE-II also shows an effect against SARS-CoV-2. It can serve as a rationale for designing a drug for patient infected with COVID-19 and associated diseases.